Chitosan/cellulose-based beads for the affinity purification of histidine-tagged proteins

被引：4
作者：
Shao, Mingcong [1 ]
Xiu, Lili [1 ]
Zhang, Haijiang [2 ]
Huang, Jianying [1 ]
Gong, Xingwen [1 ]
机构：
[1] Zhejiang Gongshang Univ, Coll Food Sci & Biotechnol, Hangzhou 310018, Zhejiang, Peoples R China
[2] Huaiyin Inst Technol, Jiangsu Key Lab Reg Resource Exploitat & Med Res, Huaian, Peoples R China
来源：
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY | 2018年 / 48卷 / 04期
基金：
中国国家自然科学基金;
关键词：
Affinity purification; chitosan; his-tagged protein; ionic liquid; reuse; CHITOSAN; CELLULOSE; ADSORPTION; EXTRACTION; ACID; IMMOBILIZATION; NANOPARTICLES; ENZYMES; LIQUID; CHITIN;
D O I：
10.1080/10826068.2018.1446154
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Chitosan/cellulose-based beads (CCBs) for the affinity purification of histidine-tagged proteins were prepared from chitosan/cellulose dissolved in ionic liquid as a solvent, and their structures were characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and thermogravimetric analysis. The affinity purification was used to separate hexahistidine-tagged (his-tagged) enhanced green fluorescent protein (EGFP) from Escherichia coli. The results showed that Zn2+-CCB exhibited more specific adsorption capacity toward the target protein compared with Ni2+-CCB and Cu2+-CCB. The maximum adsorption of EGFP was 1.84mg/g of Zn2+-CCB, with 90% purity under the optimized conditions (ionic strength (1.0M NaCl), pH (7.2) and imidazole concentration (500mM)). In addition, a regeneration method for the sorbent was further developed by washing with ethylenediaminetetraacetic acid disodium and then reimmobilizing with metal ions. This technique is an alternative method for the purification of his-tagged proteins, making the process more economical, fast, stable, and large batch.
引用
收藏
页码：352 / 360
页数：9
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